In the Netherlands 30% of all women of reproductive age are habitual smokers. One third of these women continue to smoke during pregnancy. Tobacco smoke consists of more than 3600 different compounds. One of its chief pharmacologically active ingredients is nicotine of which 60% is metabolized to cotinine. Cotinine is the best available biochemical marker of nicotine consumption because it is specific for tobacco smoke exposure and it has a relatively long mean t1/2 of 15 hours. In the present study nicotine and cotinine concentrations were measured in 25 smoking and 25 non-smoking healthy pregnant women. In all 25 non-smoking pregnant women nicotine and cotinine levels were < 10 mg/l. Light smokers (< 10 cigarettes/day) were found to have nicotine blood concentrations < 10 mg/l and cotinine levels varying between 40 and 99 mg/l. Heavy smokers (> or = 10 cigarettes/day) had nicotine concentrations < 10 mg/l, but high cotinine levels varying from 115 to 199 mg/l. Cotinine was also determined in 25 neonates of non-smoking mothers and in 34 neonates of smoking mothers. In 9 of these 34 newborns the relationship between maternal and neonatal cotinine concentrations was investigated. Cotinine levels in neonates born to non-smokers and to women who smoked less than 10 cigarettes/day were below the detection limit of 10 mg/l. Cotinine values in neonates whose mothers smoked > or = 10 cigarettes/day were significantly higher than in those whose mothers smoked < 10 cigarettes/day, but significantly lower than in their mothers. The results of this study confirm that cotinine is more useful than nicotine in discriminating non-smokers, light and heavy smokers. Cotinine concentrations were significantly lower in the neonates than in their mothers, but there was a strong positive linear relationship between maternal and neonatal cotinine concentrations.